Pharmaceutical Analysis

Glycosylation of monoclonal antibody (mAb) therapeutics is widely recognized by the regulators and the industry as a critical quality attribute (CQA). Hence, it is necessary that glycosylation is measured and adequately controlled during production. This installment reviews the various process parameters and raw material attributes that affect glycosylation, as well as the different analytical tools that are used for characterization, with greater emphasis on the chromatographic methods of analysis. Key recent advancements that have occurred in the past five years are also discussed briefly. While significant progress has been made in the monitoring of glycosylation, its real time control has yet to be demonstrated.

This fourth and last installment in the series of “Separation Science in Drug Development” provides an overview of modern practices of Quality Control in small molecule drug development including activities such as setting specifications, method validation/transfer, release and stability testing, and authoring CMC sections of regulatory filings.

The third installment in this series provides an overview of modern practices of separation science in small-molecule drug development. It highlights approaches in HPLC method development and physical/chemical characterization to support process chemistry and formulation development, and for assessment/control of the clinical trial materials. The role of the separation scientist in analytical development and salient chromatographic methodology trends are discussed.

A systematic approach for formation of sigmoidal gradient for analysis of biopharmaceutical proteins has been reviewed and discussed together with two case studies. The first application involves HPLC separation of the various product related variants of a microbial expressed biotherapeutic, granulocyte colony stimulating factor (GCSF). The second case study involves separation of the charge heterogeneity related variants for a monoclonal antibody (mAb) biotherapeutic product. In all cases, it is observed that the use of sigmoidal gradient successfully reduces the analysis time significantly (from 70 minutes to 15 minutes for GCSF and from 40 minutes to 4 minutes for the mAb) while retaining the selectivity and the resolution.

The United States Food and Drug Administration (FDA) has issued a new guidance document, Analytical Procedures and Method Validation for Drugs and Biologics. The guidance is quite general in nature. Anyone hoping for specific recommendations on topics such as which methods to use will be disappointed. Industry experts say that i really isn’t feasible for the FDA to provide detailed recommendations about analytical methods for biopharmaceuticals, however.

This installment provides an overview of high-throughput characterization techniques of drug leads to support small molecule drug discovery programs in a pharmaceutical company. A myriad of analytical chemistry techniques including separation science methodologies are used to confirm the structures and identities, quantitating the concentrations of stock solutions, and measuring key physicochemical properties of the new chemical entities (NCE). A case study is used here to illustrate the details of these applications in high-throughput characterization.

Here in part II of our series on assessing protein aggregation, we provide an overview of best practices for achieving this goal, including the importance of using a multimethod approach.Here in part II of our series on assessing protein aggregation, we provide an overview of best practices for achieving this goal, including the importance of using a multimethod approach.

This installment provides an overview of modern practices of high-throughput purification to support small-molecule drug discovery.

Benedetto Natalini of the University of Perugia, Italy, spoke to Bethany Degg of The Column about the driving forces in pharmaceutical analysis, including the importance of regulation, chirality, and miniaturization.

Experimental results and method validation data from two analytical case studies of complex small?molecule pharmaceuticals illustrates the utility and advantages of UHPLC in high-resolution separations.

Most analytical approaches used for the determination of low-molecular-weight noncovalent aggregates in protein pharmaceuticals suffer from important limitations. This first part of this two-part column series discusses those limitations.

In this installment, we discuss fluorescence-detection size exclusion chromatography (FSEC) technology and its implications in protein characterization, specifically the development and characterization of biologics.  

A discussion of the role of the gradient in separation by reversed-phase and ion-exchange HPLC, with data from two products to illustrate the key points

During the past year, LCGC examined current trends in the application of liquid chromatography (LC), and gas chromatography (GC), and related techniques in environmental, food, forensics, and pharmaceutical analysis. This article presents some developments made by separation scientists working in these application areas and offers insights into the current trends in each field.

The modern usage and specific applications of HILIC for two other major classes of analytes - glycopeptides and glycoproteins - are discussed.

Agilent Technologies

Answers to common questions about HILIC, and an overview of its application to biopharmaceutical analysis.

LCGC recently spoke with Pauline Rudd of University College, Dublin, and The National Institute for Bioprocessing Research and Training (NIBRT) in Dublin, Ireland, about her work using ultrahigh-pressure liquid chromatography (UHPLC) for the characterization of protein glycosylation.

The use of a monolithic column reduces analysis time in an on-line biopharmaceutical HPLC analysis of granulocyte colony stimulating factor.

Recent advances in sampling techniques in the pharmaceutical industry sparked significant interest in applying improvements to extraction methods for greater analyte detection and quantitation.

A method using HR-MS systems to characterize the structures of metabolites is presented. The result is a general workflow for metabolism studies in drug discovery and development.

Click here to view the complete E-Separation Solutions newsletter from August 22, 2013.

A set of extended chiral separation screens using HPLC and SFC has been tailored to meet the specific and changing needs of customers in a pharmaceutical development setting.

HPLC method development can be accelerated by using three distinct method templates of increasing complexity.

Pharmaceutical Analysis Technology Forum